Human GDA knockout HeLa cell lysate

• WB

Lab

Western blot - Human GDA knockout HeLa cell lysate (AB258431)

Lane 1 : Wild-type HeLa cell lysate 20 ug
Lane 2 : GDA knockout HeLa cell lysate 20 ug
Lane 3 : Mouse kidney cell lysate 20 ug
Lane 4 : Daudi cell lysate 20 ug
False colour image of Western blot : Anti-GDA antibody staining at 0.5 ugg/ml, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab233221 was shown to bind specifically to GDA. A band was observed at 51 kDa in wild-type HeLa cell lysates with no signal observed at this size in GDA knockout cell line ab264695 (knockout cell lysate ab258431). To generate this image, wild-type and GDA knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-GDA antibody (<a href='/en-us/products/primary-antibodies/gda-antibody-ab233221'>ab233221</a>) at 0.5 µg/mL

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human GDA knockout HeLa cell lysate (ab258431)

Lane 2:

GDA knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human GDA knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-gda-knockout-hela-cell-line-ab264695'>ab264695</a>)

Lane 3:

Mouse kidney cell lysate at 20 µg

Lane 4:

Daudi cell lysate at 20 µg

Predicted band size: 51 kDa

Observed band size: 51 kDa

false

• Sanger seq

Unknown

Sanger Sequencing - Human GDA knockout HeLa cell lysate (AB258431)

Allele-1 : 29 bp deletion in exon 3

• Sanger seq

Unknown

Sanger Sequencing - Human GDA knockout HeLa cell lysate (AB258431)

Allele-2 : 2 bp deletion in exon 3